Aim:To identify the characteristics of recombinant adenovirus modified PBMC-derived dendritic cells and their resistance to HIV-1 infection.Method:The recombinant adenovirus vector was integrated with CCR5△32,CCR5siRNA,HIV-1 poli and HIV-1 inti genes by using AdEasy system.Human PBMC from healthy donor blood was isolated for dendritic cells.The expression of CCR5△32,CCR5,CXCR4 and HIV-1 p24 in PBMC or modified cells was measured by Westernblot,p24 expression in cell lysis by ELISA and HIV-1 entrance by β-galactosidase assay.Furthermore,the T cell immunity action induced by recombinant adenovirus was measured with ELISPOT.Results:After the cells were modified by Ad-R5△32siRNA,the expression of CCR5△32 was increased while the expression of CCR5 and CXCR4 decreased.There was no adverse effect of adenoviral gene transfer on DC development.CD83 expression in the matured DC cells surface was not changed after gene transfer.The expression of p24 was remained at low levels in modified cells when challenged by HIV-1.The modified cells showed resistance to HIV-1 infection.Conclusion:Recombinant adenovirus-modified cells demonstrated good resistance to HIV-1 infection.Modification of HSC-derived immunity cells,such as DCs,may be a potent strategy to resist HIV-1 infection.
Cheng-lai Xia Ping Zhu Yan-tao Cai Guang-bin Zhu Zheng-rong Mei Hanhui-Huang Di-xian Luo Peng-ke Yan
Department of Pharmacy,The Third affiliated Hospital,Medical university of Guangzhou,Guangzhou,China Guangdong Cardiovascular Institute,Guangdong General Hospital,Guangdong Academy of Medical Sciences, Department of Dermatology and Rheumatology,Foshan Maternity & Child Heath Care Hospital,Foshan,China Department of radiology,The First Affiliated Hospital,Medical university of Guangzhou.Guangzhou,Chin Institute of Tranditional Medicine of University of South China,Chenzhou No.l People”s Hospital,Chen